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Downregulation of miR-432 activates Wnt/β-catenin signaling and promotes human hepatocellular carcinoma proliferation.

Jiang N, Chen WJ, Zhang JW, Xu C, Zeng XC, Zhang T, Li Y, Wang GY - Oncotarget (2015)

Bottom Line: Sustained cell growth and proliferation, one of the hallmarks of cancer, is considered to responsible for cancer-related deaths by disorganizing the balance of growth promotion and growth limitation.Elucidating the molecular mechanism of this abnormality in hepatocellular carcinoma carcinogenesis may improve diagnostic and therapeutic strategies for this malignancy.Furthermore, miR-432 directly targeted and suppressed multiple regulators of the Wnt/β-catenin signaling cascade, including LRP6, TRIM29 and Pygo2, which subsequently deactivated Wnt/β-catenin signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatic Surgery, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong, China.

ABSTRACT
Sustained cell growth and proliferation, one of the hallmarks of cancer, is considered to responsible for cancer-related deaths by disorganizing the balance of growth promotion and growth limitation. Aberrant activation of the Wnt/β-catenin signaling pathway leads to cell proliferation, growth and survival, and promotes the development of various human tumors, including hepatocellular carcinoma. Elucidating the molecular mechanism of this abnormality in hepatocellular carcinoma carcinogenesis may improve diagnostic and therapeutic strategies for this malignancy. Herein, we report that the expression of miR-432 was markedly downregulated in hepatocellular carcinoma cell lines and tissues, and upregulation of miR-432 inhibited, whereas downregulation of miR-432 enhanced the proliferation and tumorigenicity of hepatocellular carcinoma cells both in vitro and in vivo. Furthermore, miR-432 directly targeted and suppressed multiple regulators of the Wnt/β-catenin signaling cascade, including LRP6, TRIM29 and Pygo2, which subsequently deactivated Wnt/β-catenin signaling pathway. Finally, miR-432 expression was inversely correlated with three targets in clinical hepatocellular carcinoma samples. These results demonstrated that miR-432 functions as a tumor-suppressive miRNA by suppressing Wnt/β-catenin signaling activation and may represent a therapeutic target for hepatocellular carcinoma.

No MeSH data available.


Related in: MedlinePlus

Ectopic expression of miR-432 inhibited HCC cell proliferation(A). MTT assay revealed that miR-432 upregulation suppresses QGY-7703 and HepG2 stable cell lines at indicated times after seeding. (B). Representative micrographs (left) and quantification (right) of crystal violet-stained cell colonies. Indicated cells (0.5 × 103) were plated into six-well plates and cultured for 10 days, then stained with crystal violet (1.0%). (C). Representative micrographs (left) and quantification of colonies > 0.1 mm (right) were scored. Indicated cells (2 × 103) were suspended in soft agar and cultured for 10 days, and then colonies > 0.1 mm in diameter were counted. (D). Representative micrographs (left) and quantification of BrdU positive signaling in the cells transfected with miR-432 or Vector. (E). Flow cytometry analysis of indicated HCC cells stably expressing miR-432 or Vector. Each bar represents the mean ± SD of three independent experiments. *P < 0.05.
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Figure 2: Ectopic expression of miR-432 inhibited HCC cell proliferation(A). MTT assay revealed that miR-432 upregulation suppresses QGY-7703 and HepG2 stable cell lines at indicated times after seeding. (B). Representative micrographs (left) and quantification (right) of crystal violet-stained cell colonies. Indicated cells (0.5 × 103) were plated into six-well plates and cultured for 10 days, then stained with crystal violet (1.0%). (C). Representative micrographs (left) and quantification of colonies > 0.1 mm (right) were scored. Indicated cells (2 × 103) were suspended in soft agar and cultured for 10 days, and then colonies > 0.1 mm in diameter were counted. (D). Representative micrographs (left) and quantification of BrdU positive signaling in the cells transfected with miR-432 or Vector. (E). Flow cytometry analysis of indicated HCC cells stably expressing miR-432 or Vector. Each bar represents the mean ± SD of three independent experiments. *P < 0.05.

Mentions: To determine the effect of miR-432 on HCC progression, QGY-7703 and HepG2 cells were selected for further study (Supplemental Figure 1). As shown in Figure 2A and 2B, upregulation of miR-432 significantly decreased the growth rate of QGY-7703 and HepG2 cells, analyzed by MTT and colony formation assays. Importantly, the anchorage-independent growth ability of QGY-7703 and HepG2 cells was drastically repressed upon miR-432 overexpression cells, as indicated by the reduction in colony number and colony size on soft agar (Figure 2C), suggesting that miR-432 upregulation decreased the tumorigenicity of HCC cells in vitro.


Downregulation of miR-432 activates Wnt/β-catenin signaling and promotes human hepatocellular carcinoma proliferation.

Jiang N, Chen WJ, Zhang JW, Xu C, Zeng XC, Zhang T, Li Y, Wang GY - Oncotarget (2015)

Ectopic expression of miR-432 inhibited HCC cell proliferation(A). MTT assay revealed that miR-432 upregulation suppresses QGY-7703 and HepG2 stable cell lines at indicated times after seeding. (B). Representative micrographs (left) and quantification (right) of crystal violet-stained cell colonies. Indicated cells (0.5 × 103) were plated into six-well plates and cultured for 10 days, then stained with crystal violet (1.0%). (C). Representative micrographs (left) and quantification of colonies > 0.1 mm (right) were scored. Indicated cells (2 × 103) were suspended in soft agar and cultured for 10 days, and then colonies > 0.1 mm in diameter were counted. (D). Representative micrographs (left) and quantification of BrdU positive signaling in the cells transfected with miR-432 or Vector. (E). Flow cytometry analysis of indicated HCC cells stably expressing miR-432 or Vector. Each bar represents the mean ± SD of three independent experiments. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4480722&req=5

Figure 2: Ectopic expression of miR-432 inhibited HCC cell proliferation(A). MTT assay revealed that miR-432 upregulation suppresses QGY-7703 and HepG2 stable cell lines at indicated times after seeding. (B). Representative micrographs (left) and quantification (right) of crystal violet-stained cell colonies. Indicated cells (0.5 × 103) were plated into six-well plates and cultured for 10 days, then stained with crystal violet (1.0%). (C). Representative micrographs (left) and quantification of colonies > 0.1 mm (right) were scored. Indicated cells (2 × 103) were suspended in soft agar and cultured for 10 days, and then colonies > 0.1 mm in diameter were counted. (D). Representative micrographs (left) and quantification of BrdU positive signaling in the cells transfected with miR-432 or Vector. (E). Flow cytometry analysis of indicated HCC cells stably expressing miR-432 or Vector. Each bar represents the mean ± SD of three independent experiments. *P < 0.05.
Mentions: To determine the effect of miR-432 on HCC progression, QGY-7703 and HepG2 cells were selected for further study (Supplemental Figure 1). As shown in Figure 2A and 2B, upregulation of miR-432 significantly decreased the growth rate of QGY-7703 and HepG2 cells, analyzed by MTT and colony formation assays. Importantly, the anchorage-independent growth ability of QGY-7703 and HepG2 cells was drastically repressed upon miR-432 overexpression cells, as indicated by the reduction in colony number and colony size on soft agar (Figure 2C), suggesting that miR-432 upregulation decreased the tumorigenicity of HCC cells in vitro.

Bottom Line: Sustained cell growth and proliferation, one of the hallmarks of cancer, is considered to responsible for cancer-related deaths by disorganizing the balance of growth promotion and growth limitation.Elucidating the molecular mechanism of this abnormality in hepatocellular carcinoma carcinogenesis may improve diagnostic and therapeutic strategies for this malignancy.Furthermore, miR-432 directly targeted and suppressed multiple regulators of the Wnt/β-catenin signaling cascade, including LRP6, TRIM29 and Pygo2, which subsequently deactivated Wnt/β-catenin signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatic Surgery, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong, China.

ABSTRACT
Sustained cell growth and proliferation, one of the hallmarks of cancer, is considered to responsible for cancer-related deaths by disorganizing the balance of growth promotion and growth limitation. Aberrant activation of the Wnt/β-catenin signaling pathway leads to cell proliferation, growth and survival, and promotes the development of various human tumors, including hepatocellular carcinoma. Elucidating the molecular mechanism of this abnormality in hepatocellular carcinoma carcinogenesis may improve diagnostic and therapeutic strategies for this malignancy. Herein, we report that the expression of miR-432 was markedly downregulated in hepatocellular carcinoma cell lines and tissues, and upregulation of miR-432 inhibited, whereas downregulation of miR-432 enhanced the proliferation and tumorigenicity of hepatocellular carcinoma cells both in vitro and in vivo. Furthermore, miR-432 directly targeted and suppressed multiple regulators of the Wnt/β-catenin signaling cascade, including LRP6, TRIM29 and Pygo2, which subsequently deactivated Wnt/β-catenin signaling pathway. Finally, miR-432 expression was inversely correlated with three targets in clinical hepatocellular carcinoma samples. These results demonstrated that miR-432 functions as a tumor-suppressive miRNA by suppressing Wnt/β-catenin signaling activation and may represent a therapeutic target for hepatocellular carcinoma.

No MeSH data available.


Related in: MedlinePlus